Translational inactivation of the ndhB gene in the pSSH1 plastid transformation plasmid. The pSSH1 plasmid insert spans the first 732 bp of the ndhB gene. An additional C-G bp was introduced into codon 206 of ndhB by oligonucleotide-directed mutagenesis. The resulting plasmid was called pSSH1M. This additional nucleotide generated a diagnostic SmaI site and, in addition to a frame shift, all three stop codons (only two of which are shown). A 300-bp portion of the ndhB coding region (identical in tobacco and nightshade) adjacent to the HindIII cloning site is shown as it appears in IRA (5′–3′ direction, strand A), from position 143,798 in the tobacco plastid genome. Selected restriction enzyme sites are also shown.